This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Coordinate gene regulation is a fundamental biological process essential to all cells from the germ line to the immune system. Regulation is disrupted in disorders as diverse as autism, Crohn's disease, and Down's syndrome. X-chromosome dosage compensation is one of the best models for studying this process because thousands of genes are co-regulated. Drosophila, like mammals, increase the transcript levels from a large number of diverse genes along the single male X-chromosome precisely two-fold relative to each female X-chromosome and all other chromosomes. The Drosophila Male Specific Lethal (MSL) complex is central to dosage compensation. It has long been proposed that MSL complex first identifies high affinity sites on the male X-chromosome and then spreads along its length. However, there is no temporal data to support this hypothesis due to limitations in obtaining sufficient material for biochemistry. Therefore, we will develop an innovative cell induction system to generate sufficient material to perform chromatin immunoprecipitation followed by NextGen sequencing (ChIPseq) at many time points. We expect that this pilot project will generate the first system to define how sub-nuclear domains are formed in real-time across species and provide a strong foundation for an NIH R01.